sinomenine and Disease-Models--Animal

The rhizome of Chinese medical plant QingTeng (scientific name: Sinomenium acutum (Thunb.) Rehd. et Wils.) is widely used by traditional medical doctors for anti-inflammation and immunoregulatory in China and other Asian countries.. The purpose of this study was to evaluate the effects and possible mechanisms of sinomenine resistance against DSS-induced inflammation in vitro and in vivo.. The UC model was induced by treating female mice with 3% DSS in vivo and human colonic epithelial cells (Hcoepic) with 0.8 mg/ml DSS in vitro. The mice and Hcoepic were then treated with sinomenine. Inflammatory factors were detected using ELISA and qRT-PCR. Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 and 14-3-3θ were analyzed by bioinformatic analysis and verified by western blotting, immunofluorescent staining or immunohistochemistry.. DSS-induced Hcoepic underwent high inflammation and oxidative stress conditions, whereas sinomenine reduced the uncontrolled immune microenvironment by suppressing NF-κB signaling and targeting 14-3-3θ. Knockdown of 14-3-3θ decreased the protective effect of sinomenine against DSS-induced inflammation in vitro. Moreover, sinomenine promoted 14-3-3θ protein expression and inhibited NF-κB p65 signaling in DSS-induced mice.. These findings suggest that 14-3-3θ plays an important role in sinomenine against DSS treatment, and sinomenine could be considered a potential drug for the treatment of UC.

Topics: Animals; Colitis, Ulcerative; Dextran Sulfate; Disease Models, Animal; Humans; Inflammation; Mice; Mice, Inbred C57BL; Morphinans; NF-kappa B; Signal Transduction

Psoriasis is a chronic inflammatory skin disease. Sinomenine (SIN) has anti-inflammatory and antioxidant effects.. The objective of this study was to confirm the anti-inflammatory effects and mechanism of SIN in imiquimod (IMQ)-induced psoriasis-like mouse model and IMQ-induced differentiated human keratinocytes (HaCaT) cells.. BALB/c mice were treated with IMQ to construct a psoriasis-like mice model. PASI score and HE staining were used to observe pathology injury of skin tissue. The secretion of inflammatory factors and the oxidative stress level were detected by ELISA. HaCaT cells after induction of differentiation were treated with IMQ (100 μM) and SIN (10 μg/mL or 50 μg/mL), cell viability, the secretion of inflammatory factors, and the oxidative stress level were detected by MTT assay, ELISA, respectively. The expression of lncRNA XIST was detected by RT-qPCR. The relationship between XIST and EIF4G2 protein was detected by RNA immunoprecipitation (RIP) assay and ubiquitination experiment.. SIN significantly reduced PASI score, epidermal thickness, inflammatory response, and oxidative stress levels that increased by IMQ in vivo. SIN inhibited IMQ-induced HaCaT cell proliferation, inflammatory response, and oxidative stress levels and decreased the expression of XIST. Overexpression of XIST negated the protective effect of SIN on HaCaT cells. XIST interacted directly with EIF4G2 and regulated EIF4G2 expression via K48 ubiquitin. Knockdown of XIST reduced the half-life of EIF4G2 and decreased EIF4G2 protein stability. In addition, the E3 ubiquitin protein ligase MDM2 interacted with EIF4G2 and downregulated EIF4G2 expression. XIST reduced the interaction between MDM2 and EIF4G2, which mediated EIF4G2 K48 ubiquitination. Overexpression of XIST negated the protective effect of SIN on the inflammation of HaCaT cells through activating the NF-κB signaling pathway, while NF-κB pathway inhibitor PDTC reversed this result.. SIN had a protective effect on psoriasis and could inhibit HaCaT cell proliferation and inflammatory response via XIST/MDM2/EIF4G2/NF-κB axis.

Topics: Animals; Anti-Inflammatory Agents; Cell Proliferation; Dermatitis; Disease Models, Animal; Humans; Imiquimod; Keratinocytes; Mice; Mice, Inbred BALB C; NF-kappa B; Psoriasis; RNA, Long Noncoding; Skin Diseases

Sepsis is a systemic inflammatory response syndrome caused by a host's immune response to infection. Acute lung injury (ALI) is one of the most common complications of sepsis with high mortality and morbidity. Recent evidence demonstrated that the 'gut-lung axis' was related to the progression of septic acute lung injury, which regarded gut microbiota and intestinal barrier as two critical factors correlated with acute lung injury. Sinomenine is an isoquinoline alkaloid component extracted from Sinomenium acutum Rehd，et Wils, which has been already reported to have significant anti-inflammatory, immunosuppressive, and anti-arthritis properties. In this research, we observed that sinomenine could repair the lung injury and alleviate inflammatory response induced by cecum ligation and puncture (CLP). Illumine sequencing of 16S rDNA revealed that sinomenine could improve the richness of gut microbiota and modulate the composition of intestinal flora in cecum ligation and puncture mice. Meanwhile, sinomenine could reduce the colon pathological damage and improve the intestine barrier integrity in cecum ligation and puncture mice. We also found that the molecular mechanism of sinomenine's protective effect on intestinal tract was related to the activation of aryl hydrocarbon receptor/nuclear factor erythroid-2 related factor 2（Nrf2）pathway both in vivo and vitro experiments. Collectively, the prevention of septic acute lung injury by sinomenine might be mediated by modulating gut microbiota and restoring intestinal barrier via aryl hydrocarbon receptor/Nrf2-dependent pathway.

Topics: Acute Lung Injury; Animals; Anti-Inflammatory Agents; Basic Helix-Loop-Helix Transcription Factors; Cecum; Cell Line, Tumor; Cytokines; Disease Models, Animal; Gastrointestinal Microbiome; Gastrointestinal Tract; Homeostasis; Humans; Inflammation; Ligation; Male; Mice, Inbred ICR; Morphinans; NF-E2-Related Factor 2; Permeability; Protective Agents; Punctures; Receptors, Aryl Hydrocarbon

Topics: Airway Remodeling; Animals; Anti-Asthmatic Agents; Asthma; Cell Line; Cell Movement; Cell Proliferation; Disease Models, Animal; Epithelial Cells; Epithelial-Mesenchymal Transition; Female; Humans; Interleukin-4; Lung; Mice, Inbred BALB C; Morphinans; Ovalbumin; Signal Transduction; Smad3 Protein; Transforming Growth Factor beta1

Topics: Animals; Astrocytes; Body Weight; Cytokines; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Encephalomyelitis, Autoimmune, Experimental; Female; Inflammasomes; Mice; Mice, Inbred C57BL; Microglia; Morphinans; Myelin-Oligodendrocyte Glycoprotein; NLR Family, Pyrin Domain-Containing 3 Protein; Peptide Fragments; Pyroptosis; Random Allocation; Specific Pathogen-Free Organisms; Spinal Cord

BACKGROUND This study aims to explore the effect of Sinomenine (SIN) on pregnancy outcomes of recurrent spontaneous abortion (RSA) in a mouse model. MATERIAL AND METHODS Thirty female CBA/J mice were allocated into 3 groups randomly, then mated with BALB/c mice (CBA/J×BALB/c) as normal-pregnancy group (n=10), or mated with DBA/2 mice (CBA/J×DBA/2) as RSA model (n=10), or CBA/J×DBA/2 mice treated with SIN as RSA+SIN group (n=10). The number of surviving and reabsorbed embryos in each group were counted on day 13.5 of gestation. The mouse serum was collected to determine the levels of interferon-γ (IFN)-γ and IL-4 by ELISA. Immunohistochemistry, qRT-PCR and immunoblotting were used to determine the location, mRNA and protein expressions of IFN-γ, IL-4, T-bet and GATA3 in the decidual and placental tissue. RESULTS In the RSA group, the amount of reabsorbed embryo was significantly higher than that in the normal-pregnancy group. However, SIN treatment showed a rescue effect on spontaneous abortion in RSA mice. IFN-γ, IL-4, T-bet, and GATA3 were all expressed in placental tissues and mainly located in the cytoplasm. The RSA group demonstrated higher expression levels of IFN-γ and T-bet than in the RSA+SIN and normal-pregnancy groups. Although RSA and RSA+SIN groups showed lower expression levels of IL-4 and GATA3 than in the normal-pregnancy group, there was no significant difference between RSA and RSA+SIN groups regarding IL-4 and GATA expression levels. CONCLUSIONS SIN treatment demonstrates a therapeutic effect on spontaneous abortion in RSA mice, possibly through regulating the balance of Th1/Th2 in maternal circulation and decidual tissues.

Topics: Abortion, Spontaneous; Animals; Decidua; Disease Models, Animal; Embryo, Mammalian; Female; GATA3 Transcription Factor; Interferon-gamma; Interleukin-4; Male; Mice; Morphinans; Placenta; Pregnancy; T-Box Domain Proteins; Th1-Th2 Balance

The objective of study was to investigate the inhibitory effect of sinomenine on neuropathic pain on dorsal root ganglia (DRG). The DRG cell line and spinal nerve ligation (SNL) model were used in this study. The effect of sinomenine on the cell viability was examined by MTT assay. The expression of p38 MAPK, NF-κB, c-fos, SP and TNF-α was detected by using immunofluorescence and immunohistochemistry assay. We also assessed the level of p-CaMKII, COX-2, p-CREB, IL-17A, TLR4 and IL-1β via western blotting and RT-qPCR. Compared to the controls, sinomenine showed a protective effect on TNF-α-induced apoptosis on DRG cells in a dose-dependent manner, with an increase of cell viability and a decrease of reactive oxygen species level as well as LDH release. Parallelly, sinomenine treatment significantly reduced the expression of various factors related to stress and inflammation, including p38 MAPK, NF-κB, c-fos, p-CAMKII, COX-2, p-CREB, TLR4 and IL-17A in DRG cells in vitro. Furthermore, we found that administration of sinomenine significantly reduced mechanical withdrawal threshold and thermal withdrawal latency and inhibited the inflammation and activation of p38 signaling in SNL rats. It is noting that combined therapy of sinomenine and pulsed radiofrequency exhibited higher efficacy of dorsal root ganglia inflammation than single treatment as well as the combination of oxycodone and pulsed radiofrequency. Sinomenine inhibited the apoptosis of DRG cell by regulating p38 MAPK/CREB signalling pathway, which provides evidence to alleviate neuropathic pain in clinic.

Topics: Analgesics; Analgesics, Opioid; Animals; Anti-Inflammatory Agents; Apoptosis; Behavior, Animal; Cell Line; Combined Modality Therapy; Cyclic AMP Response Element-Binding Protein; Disease Models, Animal; Ganglia, Spinal; Inflammation; Inflammation Mediators; Male; Morphinans; Neuralgia; Oxycodone; p38 Mitogen-Activated Protein Kinases; Pain Threshold; Pulsed Radiofrequency Treatment; Rats, Sprague-Dawley; Signal Transduction

When Zika virus emerged as a public health emergency there were no drugs or vaccines approved for its prevention or treatment. We used a high-throughput screen for Zika virus protease inhibitors to identify several inhibitors of Zika virus infection. We expressed the NS2B-NS3 Zika virus protease and conducted a biochemical screen for small-molecule inhibitors. A quantitative structure-activity relationship model was employed to virtually screen ∼138,000 compounds, which increased the identification of active compounds, while decreasing screening time and resources. Candidate inhibitors were validated in several viral infection assays. Small molecules with favorable clinical profiles, especially the five-lipoxygenase-activating protein inhibitor, MK-591, inhibited the Zika virus protease and infection in neural stem cells. Members of the tetracycline family of antibiotics were more potent inhibitors of Zika virus infection than the protease, suggesting they may have multiple mechanisms of action. The most potent tetracycline, methacycline, reduced the amount of Zika virus present in the brain and the severity of Zika virus-induced motor deficits in an immunocompetent mouse model. As Food and Drug Administration-approved drugs, the tetracyclines could be quickly translated to the clinic. The compounds identified through our screening paradigm have the potential to be used as prophylactics for patients traveling to endemic regions or for the treatment of the neurological complications of Zika virus infection.

Topics: Animals; Antiviral Agents; Artificial Intelligence; Chlorocebus aethiops; Disease Models, Animal; Drug Evaluation, Preclinical; High-Throughput Screening Assays; Immunocompetence; Inhibitory Concentration 50; Methacycline; Mice, Inbred C57BL; Protease Inhibitors; Quantitative Structure-Activity Relationship; Small Molecule Libraries; Vero Cells; Zika Virus; Zika Virus Infection

This study is designed to investigate the effects and mechanisms of sinomenine (Sin) in stress load-induced heart failure in mice.. We used aortic constriction (AB) to cause pressure overload as our heart failure model. Sin was received in mice as the treatment group. Cardiac function and structural changes were detected using echocardiography. Heart-lung mass ratios were measured. The serum levels of IL-10 and IL-17 proteins were detected by using ELISA, cardiac hypertrophy markers atrial natriuretic peptide (ANP), myocardial I and III collagen mRNA levels were detected by RT-PCR. Myocardial type I and III collagen protein levels were detected by Western blotting.. Sin significantly improved stress load-induced heart failure (P < 0.05), reduced the heart-lung mass ratio, ANP, collagen-I and -III mRNA and protein levels (P < 0.05); Sin can enhance the ratio of IL-10/IL-17.. Sin may be a promising drug target to improve heart failure. Its role is related to reduce serum ANP levels, inhibit the mRNA and protein level of type I and III collagen and enhance the ratio of IL-10/IL-17.

Topics: Animals; Atrial Natriuretic Factor; Cardiotonic Agents; Collagen Type I; Collagen Type III; Disease Models, Animal; Heart Failure; Interleukin-10; Interleukin-17; Male; Mice; Mice, Inbred C57BL; Morphinans

Plasma cell mastitis (PCM) is a special form of mastitis characterized by periductal inflammation and large-scale plasma cell infiltration. At present, the recurrence rate of PCM after excision is quite high, making PCM a major problem for mammary surgeons. However, no effective drug exists for the treatment of PCM. Numerous studies have demonstrated that Sinomenine hydrochloride (SH) has potent anti-inflammatory and immunoregulatory properties. However, the efficacy and the underlying mechanisms of SH in the treatment of PCM remain unclear. In the present study, we first investigated the therapeutic effects of SH in the PCM mouse model and clarified the possible mechanisms. We found that the levels of plasmocytes and lymphocytes infiltration were alleviated significantly in the 100 mg/kg SH group compared to the control group. In addition, few CD138+ plasma cells were found in the mammary glands of the 100 mg/kg SH group. The levels of Bcl-2 in the 100 mg/kg SH group were dramatically decreased compared with those in the saline group. Mechanistically, we demonstrated that SH inhibited the progression of PCM mainly through downregulating IL-6/JAK2/STAT3 levels. Collectively, our results suggested that SH could inhibit the progression of PCM by suppressing IL-6/JAK2/STAT3 cascades and ultimately achieve a therapeutic effect in PCM. This study provides theoretical support for the clinical application of SH in the treatment of PCM.

Topics: Animals; Anti-Inflammatory Agents; Disease Models, Animal; Disease Progression; Female; Humans; Interleukin-6; Janus Kinase 2; Mammary Glands, Animal; Mastitis; Mice; Morphinans; Plasma Cells; Signal Transduction; STAT3 Transcription Factor; Tyrphostins

The conventional medications are still facing a huge challenge for the treatment of rheumatoid arthritis (RA). Thus, looking for an effective therapy of RA has became an urgent issue nowadays. In this study, a novel thermosensitive liposome loaded with sinomenine hydrochloride (SIN-TSL) was developed by a pH gradient method. The SIN-TSL had a mean particle size of around 100 nm, and an high entrapment efficiency and drug loading capacity. The results also suggested that SIN-TSL had a thermosensitive drug release behaviour, with the drug release rate at 43 °C was much faster than the one at 37 °C. The SIN-TSL could be effectively taken up by lipopolysaccharide-activated HUVECs, without any cytotoxicity was observed. In addition, both in vitro and in vivo studies indicated that the SIN-TSL combined with microwave hyperthermia exhibited superior anti-rheumatoid arthritis effect. Overall, these results suggest that SIN-loaded thermosensitive liposomes combined with microwave hyperthermia could provide an optional strategy for alleviating the clinical symptoms of RA.

Topics: 1,2-Dipalmitoylphosphatidylcholine; Animals; Antirheumatic Agents; Arthritis, Rheumatoid; Cells, Cultured; Cholesterol; Combined Modality Therapy; Cytokines; Disease Models, Animal; Drug Compounding; Drug Liberation; Female; Human Umbilical Vein Endothelial Cells; Humans; Hyperthermia, Induced; Joints; Lipids; Liposomes; Microwaves; Morphinans; Particle Size; Rats, Wistar; Solubility

Traumatic brain injury (TBI) is a significant medical problem with limited treatment options and is one of the main causes of life-long disability. Neuroinflammation orchestrated by activated microglia/macrophages at the site of injury plays a critical role in the onset of many pathological events following TBI, leading to blood brain barrier (BBB) dysfunction, neuronal damage and long term neuronal and behavioral deficits. Current treatment involves intravenous administration of anti-inflammatory drugs which have limited clinical outcomes only when dosed within the early time window after injury. Hence there is an urgent need to develop improved drug delivery systems which have potential to cross impaired BBB, target and deliver drugs selectively to activated microglia/macrophages at the sites of injury, and suppress the detrimental effects of acute inflammation. In this study, we have used Sinomenine (Sino), a potent anti-inflammatory and antioxidant drug conjugated to hydroxyl terminated generation-4 PAMAM dendrimer (D-Sino) as a potential therapy for attenuating early inflammation in TBI. D-Sino conjugates were synthesized using highly robust copper-catalyzed click reaction with high purity. D-Sino conjugates enhanced the intracellular availability of Sino due to their rapid cellular uptake, significantly attenuated early/acute inflammation by suppressing pro-inflammatory cytokines (TNF-α, IL-1β, CCL-3 and IL-6), and reduced oxidative stress (iNOS and NO) in LPS activated murine macrophages (RAW 264.7) by inhibiting NF-κB activation and its nuclear translocation (the root cause for inflammation inception) significantly more as compared to the free drug. Upon systemic administration in a rabbit model of pediatric TBI, D-Sino conjugates specifically targeted activated microglia/macrophages at the site of injury in the brain. Single dose of D-Sino attenuated inflammation in the injured brain areas by suppressing inflammatory cytokines expression whereas free Sino treatment did not demonstrate a significant effect. Together, these results suggest that D-Sino conjugate may open up new avenues for increasing the therapeutic window in the treatment of early inflammation and for improving the efficacy of the drug in TBI. Moreover, this treatment can work in conjunction with current clinical practices such as therapeutic hypothermia and pharmacologically induced coma for many indications associated with TBI, where acute inflammation plays a critical role in di

Topics: Animals; Brain Injuries, Traumatic; Child; Dendrimers; Disease Models, Animal; Humans; Mice; Microglia; Morphinans; Rabbits

Oxidative stress, inflammation and pyroptosis are three of the most important mechanisms in the pathophysiology of temporal lobe epilepsy (TLE). Most people with TLE are refractory to the existing drugs. Sinomenine has shown neuroprotective effects through counteracting oxidative stress, inflammation and pyroptosis. In this study, we evaluated the effect of sinomenine on seizure behavior, oxidative stress, inflammation and pyroptosis markers in addition to its neuroprotective potential in intrahippocampal kainate-induced rat model of TLE. For this purpose, male rats (n = 60) were randomly divided into five groups, i.e., sham, kainate (lesion) with an intrahippocampal injection of kainate, kainate groups receiving sinomenine at doses of 30 or 50 mg/kg, and kainate group receiving valproic acid at a dose of 200 mg/kg (as the positive control). Our obtained data showed that sinomenine administration at a dose of 50 mg/kg can significantly decreases severity of seizures and incidence of status epilepticus (SE), hippocampal aberrant MFS and DNA fragmentation and prevents reduction of neuronal density. It also significantly restored level of ROS, MDA, HO-1 and SOD but its effect on GSH level was not significant. Additionally, sinomenine at a dose of 50 mg/kg partially counteracted the increase of NF-κB, TLR 4, TNFα, GFAP and caspase 1. These results suggest that sinomenine has anticonvulsant and neuroprotective effects by reducing hippocampal oxidative stress, inflammation, pyroptosis and apoptosis in intrahippocampal kainate model of TLE.

Topics: Animals; Anticonvulsants; Disease Models, Animal; Epilepsy, Temporal Lobe; Hippocampus; Inflammation; Kainic Acid; Male; Morphinans; Neuroprotective Agents; Oxidative Stress; Pyroptosis; Rats; Reactive Oxygen Species

To investigate the effect of topical sinomenine (SIN) on ocular surface damage in experimental dry eye in mice.. Experimental dry eye was created using scopolamine hydrobromide in female C57BL/6 mice. Eye drops consisting of 0.05%, or 0.1% SIN or phosphate-buffered saline (PBS) were applied to the experimental dry eye in mice. Tear product and corneal staining scores were measured at 7 and 14 days after treatment. Interleukin (IL)-1β and tumour necrosis factor (TNF)-α levels in the SIN groups at 14 days after treatment were compared with those of other groups.. Mice treated with 0.05% or 0.1% SIN showed a significant improvement in tear product and corneal irregularity compared to the control and PBS-treated groups. A significant decrease in the levels of IL-1βand TNF-α was observed in the 0.05% and 0.01% SIN-treated groups.. Topical SIN eye drop application can effectively improve clinical signs and decrease inflammation in the ocular surface, and alleviate ocular surface damage in dry eye.

Topics: Animals; Cornea; Disease Models, Animal; Dry Eye Syndromes; Female; Interleukin-1beta; Mice, Inbred C57BL; Morphinans; Ophthalmic Solutions; Scopolamine; Tears; Tumor Necrosis Factor-alpha

The objective of the present study is to investigate the inhibitory effects of sinomenine (SIN) on angiogenesis in a collagen-induced arthritis (CIA) mouse model.. Arthritis assessments for all mice were recorded. The histopathological assessments were performed following haematoxylin and eosin (HE) staining. Immunohistochemistry and enzyme-linked immunosorbent assay (ELISA) analyses were used to detect the expression of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and angiopoietin 1 (ANG-1) in the serum and in the membrane. Immunohistochemistry was employed to detect the synovium microvessel density (MVD).. Compared with the CIA model group, SIN significantly ameliorated swelling and erythema extension, decreased the arthritis index, reduced inflammation, cartilage damage and bone erosion, and lessened the number of CD31 positive cells on the synovium. Moreover, the levels of HIF-1α, VEGF and ANG-1 in the synovium and in the peripheral serum were increased in the untreated CIA model group but were significantly reduced in the 30 mg/kg, 100 mg/kg and 300 mg/kg SIN treatment groups.. SIN could mitigate CIA by inhibiting angiogenesis, and the mechanism may associate with the HIF-1α-VEGF-ANG-1 axis. Additionally, our study provides a referable experimental basis for the use of SIN for the treatment of rheumatoid arthritis.

Topics: Angiogenesis Inhibitors; Angiopoietin-1; Animals; Antirheumatic Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Collagen; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Hypoxia-Inducible Factor 1, alpha Subunit; Mice; Mice, Inbred C57BL; Morphinans; Neovascularization, Pathologic; Synovial Membrane; Vascular Endothelial Growth Factor A

Management of chronic pain is restricted by the lack of effective tools. This study evaluated the efficacies of sinomenine combined gabapentin or ligustrazine hydrochloride in treating peripheral and central chronic neuropathic pain. The study was conducted in mice with photochemically induced sciatic nerve injury, and in rats with photochemically induced spinal cord injury. For assessing the effectiveness of combined therapy, sinomenine, gabapentin or ligustrazine hydrochloride was injected intraperitoneally (i.p.), and pain behavioral tests were performed. At sub-effective dosages, pre-administration of sinomenine (for 60 min) plus gabapentin or ligustrazine hydrochloride, generated significant anti-allodynic effects in mice or rats with peripheral or central neuropathic pain. However, these effects were abolished when gabapentin or ligustrazine hydrochloride was pre-administered, and then sinomenine was given 60 min later. The combined efficacies of sinomenine and gabapentin or ligustrazine hydrochloride, cannot be blocked or reversed by the naloxone, suggesting the underlying mechanism is not mediated by opioid receptors. Moreover, following repeated treatments, sinomenine and gabapentin combination increased the baseline mechanical threshold, while generating prolonged analgesia, without introducing notable side effects. Sinomenine can enhance the efficacy of gabapentin or ligustrazine hydrochloride in rodent models of peripheral or central neuropathic pain, without introducing tolerance or other notable side effects. Findings of current study suggest that combing sinomenine and gabapentin or ligustrazine hydrochloride could be highly beneficial in neuropathic pain therapies.

Topics: Analgesics; Animals; Disease Models, Animal; Drug Synergism; Gabapentin; Male; Mice; Mice, Inbred C57BL; Morphinans; Neuralgia; Peripheral Nerve Injuries; Pyrazines; Spinal Cord Injuries

Sinomenine is a bioactive alkaloid extracted from Sinomenium acutum. Here, we investigated the antidepressant effects of sinomenine in mice. The antidepressant actions of sinomenine were first examined in the forced-swim test and the tail-suspension test, and then assessed in the chronic social defeat stress (CSDS) model of depression. Changes in the brain-derived neurotrophic factor (BDNF) signaling pathway after CSDS and sinomenine treatment were also investigated. A tryptophan hydroxylase inhibitor and a BDNF signaling inhibitor were also used to determine the pharmacological mechanisms of sinomenine. It was found that sinomenine induced antidepressant-like effects in the forced-swim test and tail-suspension test without affecting the locomotor activity of mice. Sinomenine also prevented the CSDS-induced depressive-like symptoms. Moreover, sinomenine fully restored the CSDS-induced decrease in the hippocampal BDNF signaling pathway, whereas a BDNF signaling inhibitor, but not a tryptophan hydroxylase inhibitor, blocked the antidepressant effects of sinomenine. In conclusion, sinomenine exerts antidepressant effects in mice by promoting the hippocampal BDNF signaling pathway.

Topics: Animals; Antidepressive Agents; Brain-Derived Neurotrophic Factor; Depression; Depressive Disorder; Disease Models, Animal; Hippocampus; Male; Mice; Mice, Inbred C57BL; Morphinans; Neurogenesis; Serotonin; Signal Transduction; Stress, Psychological

Fibroblast like synoviocyte (FLS) is a crucial in the pathogenesis of rheumatoid arthritis (RA), and involved in inflammation and joint destruction. Sinomenine (SIN), an alkaloid derived from the plant Sinomenium acutum, has anti-inflammatory and analgesic effect and been used for RA treatment in China. Alpha 7 nicotinic acetylcholine receptors (α7nAChR), as the key receptor in cholinergic anti-inflammatory pathway (CAP) to inhibit inflammation, has been detected in RA patients synovium, but its role is still unclear. Here we investigated the association between the aggressive proliferation of FLS and α7nAChR expression and the effect of sinomenine. FLS was isolated from synovial tissues of adjuvant-induced-arthritis (AIA) rat. Tumor necrosis factor(TNF)-α was used to induce the aggressive proliferation of FLS. MTT assay was applied to evaluate the proliferation of FLS. The messenger RNA (mRNA) and protein levels of α7nAChR and early growth response gene-1 (Egr-1) were measured. The results showed that TNF-α induced FLS proliferation in vitro (P < .01) and increased the phosphorylation of ERK1/2 and the expression of Egr-1 and α7nAChR (P < .05 or P < .01). U0126, the inhibitor of ERK1/2 inhibited α7nAChR expression and FLS proliferation significantly (P < .05 or P < .01). Specific short interference RNA(siRNA) of α7nAChR decreased α7nAChR expression and inhibited FLS proliferation as well. SIN inhibited the proliferation of FLS and decreased the phosphorylation of ERK1/2, and the expression of Egr-1 and α7nAChR induced by TNF-α (P < .05). In conclusion, the expression of α7nAChR involved in the aggressive proliferation of FLS induced by TNF-α and was regulated by ERK/Egr-1 signal pathway. SIN inhibited FLS proliferation and α7nAChR expression through inhibiting ERK/Egr-1 signal pathway, this may contribute to the anti-inflammatory and anti-arthritic effect of SIN.

Topics: alpha7 Nicotinic Acetylcholine Receptor; Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Cell Proliferation; Cells, Cultured; Disease Models, Animal; Early Growth Response Protein 1; Extracellular Signal-Regulated MAP Kinases; Gene Expression Regulation; Humans; Male; Morphinans; Rats; Rats, Sprague-Dawley; RNA, Small Interfering; Signal Transduction; Sinomenium; Synoviocytes

Sinomenine (SIN) has been shown to have protective effects against brain damage following traumatic brain injury (TBI). However, the mechanisms and its role in these effects remain unclear. This study was conducted to investigate the potential mechanisms of the protective effects of SIN.. The weight-drop model of TBI in Institute of Cancer Research (ICR) mice were treated with SIN or a vehicle via intraperitoneal administration 30 min after TBI. All mice were euthanized 24 h after TBI and after neurological scoring, a series of tests were performed, including brain water content and neuronal cell death in the cerebral cortex.. The level of cytochrome. SIN protected neuronal cells by protecting them against apoptosis via mechanisms that involve the mitochondria following TBI.

Topics: Animals; Apoptosis; Apoptosis Regulatory Proteins; Brain Edema; Brain Injuries, Traumatic; Cerebral Cortex; Cytochromes c; Cytoprotection; Disease Models, Animal; Dose-Response Relationship, Drug; Glutathione Peroxidase; Male; Malondialdehyde; Mice, Inbred ICR; Mitochondria; Morphinans; Nerve Degeneration; Neurons; Neuroprotective Agents; Oxidative Stress; Signal Transduction; Superoxide Dismutase-1

Sinomenine, a major bioactive ingredient isolated from traditional Chinese medicine Sinomenium acutum, has been reported to have analgesic effects in various pain animal models. N-demethylsinomenine, the N-demethylated product of sinomenine, has been identified to be the major metabolite of sinomenine and is also a natural component extracted from Sinomenium acutum. This study examined the anti-allodynic effects of N-demethylsinomenine in a mouse model of postoperative pain. A significant and sustained mechanical allodynia that lasted for 4 days was induced by making a surgical incision on the right hind paw in mice. Acute treatment with N-demethylsinomenine (10-40 mg/kg, s.c.) relieved the mechanical allodynia in a dose-dependent manner. Although there was no difference in maximal analgesic effect between N-demethylsinomenine (40 mg/kg, s.c.) and sinomenine (40 mg/kg, s.c.), the onset of action of N-demethylsinomenine was quicker than sinomenine. Repeated treatment with N-demethylsinomenine (10-40 mg/kg/day, s.c.) also dose-dependently exerted sustained antinociception against postoperative allodynia and did not produce analgesic tolerance and carry-over effect. The anti-allodynia induced by N-demethylsinomenine (40 mg/kg, s.c.) was attenuated by bicuculline, a selective γ-aminobutyric acid type A (GABA

Topics: Analgesics; Animals; Bicuculline; Disease Models, Animal; Dose-Response Relationship, Drug; GABA-A Receptor Antagonists; Hyperalgesia; Male; Mice; Mice, Inbred ICR; Morphinans; Pain, Postoperative; Receptors, GABA-A

Sinomenine, a bioactive alkaloid isolated from the traditional Chinese herb Sinomenium acutum, possesses antiinflammatory, antinociceptive, antifibrotic, and antitumorigenic properties. In this work, we sought to explore the biological effects of sinomenine on glioma cells. It was found that sinomenine caused a concentration-dependent inhibition of viability in both U87 and U251 glioma cells. Sinomenine at 16 μmol/L caused 55% to 60% reduction in the proliferation of U87 and U251 cells. Moreover, sinomenine treatment induced a G0/G1 cell cycle arrest and apoptosis. Mechanistically, sinomenine promoted p53 expression and acetylation and reduced the expression of sirtuin 1. Ectopic expression of sirtuin 1 significantly prevented sinomenine-induced p53 acetylation and growth suppression in glioma cells. Moreover, sinomenine inhibited the growth of U87 xenograft tumors in vivo and raised the p53 protein expression. Collectively, sinomenine shows antiproliferative effects against glioma cells which is mediated through downregulation of sirtuin 1 and induction of p53 activity.

Topics: Acetylation; Animals; Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cell Proliferation; Cell Survival; Disease Models, Animal; G1 Phase Cell Cycle Checkpoints; Glioma; Humans; Mice; Morphinans; Sirtuin 1; Tumor Suppressor Protein p53; Xenograft Model Antitumor Assays

Epilepsy is a common neurological disorder and is not well controlled by available antiepileptic drugs (AEDs). Inflammation is considered to be a critical factor in the pathophysiology of epilepsy. Sinomenine (SN), a bioactive alkaloid with anti-inflammatory effect, exerts neuroprotective activity in many nervous system diseases. However, little is known about the effect of SN on epilepsy.. The chronic epilepsy model was established by pentylenetetrazole (PTZ) kindling. Morris water maze (MWM) was used to test spatial learning and memory ability. H.E. staining and Hoechst 33258 staining were used to evaluate hippocampal neuronal damage. The expression of nucleotide oligomerization domain (NOD)-like receptor protein 1 (NLRP1) inflammasome complexes and the level of inflammatory cytokines were determined by western blot, quantitative real-time PCR and enzyme-linked immunosorbent assay (ELISA) kits.. SN (20, 40, and 80 mg/kg) dose-dependently disrupts the kindling acquisition process, which decreases the seizure scores and the incidence of fully kindling. SN also increases the latency of seizure and decreases the duration of seizure in fully kindled rats. In addition, different doses of SN block the hippocampal neuronal damage and minimize the impairment of spatial learning and memory in PTZ kindled rats. Finally, PTZ kindling increases the expression of NLRP1 inflammasome complexes and the levels of inflammatory cytokines IL-1β, IL-18, IL-6, and TNF-α, which are all attenuated by SN in a dose- dependent manner.. SN exerts anticonvulsant and neuroprotective activity in PTZ kindling model of epilepsy. Disrupting the kindling acquisition, which inhibits NLRP1 inflammasome-mediated inflammatory process, might be involved in its effects.

Topics: Analysis of Variance; Animals; Anticonvulsants; CARD Signaling Adaptor Proteins; Caspase 1; Convulsants; Cytokines; Disease Models, Animal; Epilepsy; Kindling, Neurologic; Male; Maze Learning; Morphinans; Nerve Tissue Proteins; Pentylenetetrazole; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Sprague-Dawley; RNA, Messenger

BACKGROUND This study aimed to investigate the effect and underlying molecular mechanism of sinomenine (SIN) on ankylosing spondylitis (AS). MATERIAL AND METHODS To study the potential role of SIN in the pathogenesis of AS, an AS mouse model was established and mice were treated with different concentrations of SIN (10, 30, and 50 mg/kg, administered intraperitoneally). Markers of inflammation and oxidative stress were determined by ELISA assay. Western blot analysis and qRT-PCR were used to quantify the levels of related proteins and gene mRNA expression. RESULTS The results suggest that AS mice has higher levels of TNF-α, IL-1β, and IL-6 (p<0.01 for all), and lower levels of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-PX) (p<0.01 for all). SIN treatment reduced the level of TNF-α, IL-1β, and IL-6 in a dose-dependent manner, and the levels of SOD, CAT, and GSH-PX were dose-dependently increased (p<0.05 for all). The results also revealed that NF-κBp65 expression decreased, while the level of IkB increased, in a dose-dependent manner, after SIN treatment in AS mice (p<0.05 for all). The level of p-p38 was dose-dependently reduced in AS mice by SIN treatment (p<0.05). Moreover, SIN inhibited Cox-2 expression in AS mice in a dose-dependent manner (p<0.05). CONCLUSIONS SIN has a beneficial role in AS through suppressing inflammatory mediators and by down-regulating oxidative stress via inhibiting the MAPKp38/NF-kB pathway and Cox-2 expression.

Topics: Animals; Antioxidants; Catalase; Disease Models, Animal; Glutathione Peroxidase; Inflammation; Interleukin-1beta; Interleukin-6; Mice; Mice, Inbred BALB C; Morphinans; NF-kappa B; Oxidative Stress; Signal Transduction; Spondylitis, Ankylosing; Superoxide Dismutase; Tumor Necrosis Factor-alpha

BACKGROUND Sinomenine (SIN) is an extract of the Chinese medicinal herb Sinomenium acutum; it has various pharmacological properties, including immunosuppression and anti-inflammation. The present study aimed to investigate whether SIN has an anti-depressant-like effect in a mouse model of depression induced by chronic unpredictable mild stress (CUMS), and to explore the underlying molecular mechanisms. MATERIAL AND METHODS A mouse model of depression was established and treated with different concentrations of SIN (30, 100, or 300 mg/kg). Then, behavioral tests, including sucrose preference test (SPT), forced swimming test (FST), and the tail suspension test (TST), were performed. The levels of norepinephrine (NE), 5-hydroxytryptamine (5-HT), and proinflammatory cytokines (interleukin-1β [IL-1β] interleukin-6 [IL-6], and tumor necrosis factor-α [TNF-α]) in the hippocampus of mice were detected by ELISA assay. The levels of p-p38, p-p65, NLRP3, ASC, and caspase-1 were measured by Western blot or/and qRT-PCR. RESULTS The results showed that SIN significantly relieved CUMSinduced depressive-like behaviors. Compared with the model mice, SIN treatment significantly increased the sucrose preference of the mice, and the immobility time in the forced swimming and the tail suspension test were shortened. In addition, SIN decreased CUMS-induced reduction in the concentrations of NE and 5-HT in the hippocampus of mice. SIN reduced CUMS-induced increases in the levels of IL-1β, IL-6, and TNF-α in the hippocampus of mice. Furthermore, activation of the p38MAPK-NF-κB pathway and the nucleotide binding and oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome induced by CUMS were inhibited by SIN treatment. CONCLUSIONS In conclusion, our results indicate the antidepressantlike effects of SIN on chronic unpredictable mild stress-induced depression in a mouse model.

Topics: Animals; Antidepressive Agents; Behavior, Animal; China; Depression; Depressive Disorder; Disease Models, Animal; Hippocampus; Inflammation; Interleukin-1beta; Interleukin-6; Male; Mice; Mice, Inbred ICR; Morphinans; Norepinephrine; Serotonin; Stress, Psychological; Tumor Necrosis Factor-alpha

Sinomenium acutum has been used in traditional medicine to treat a painful disease such as rheumatic arthritis and neuralgia. Sinomenine, which is a main bioactive ingredient in Sinomenium acutum, has been reported to have an analgesic effect in diverse pain animal models. However little is known about the detailed mechanisms underlying peripheral analgesic effect of sinomenine. In the present study, we aimed to elucidate its cellular mechanism by using formalin-induced acute inflammatory pain model in mice. We found that intraperitoneal (i.p.) administration of sinomenine (50mg/kg) suppressed formalin-induced paw licking behavior in both the first and the second phase. Formalin-induced c-Fos protein expression was also suppressed by sinomenine (50mg/kg i.p.) in the superficial dorsal horn of spinal cord. Whole-cell patch-clamp recordings from small-sized dorsal root ganglion (DRG) neurons revealed that sinomenine reversibly increased the spike threshold and the threshold current intensity for evoking a single spike and decreased firing frequency of action potentials evoked in response to a long current pulse. Voltage-gated sodium currents (I

Topics: Animals; Antirheumatic Agents; Cells, Cultured; Disease Models, Animal; Dose-Response Relationship, Drug; Formaldehyde; Ganglia, Spinal; Inflammation; Male; Mice; Mice, Inbred C57BL; Morphinans; Neuralgia; Pain; Pain Measurement; Patch-Clamp Techniques; Proto-Oncogene Proteins c-fos; Sensory Receptor Cells; Sodium; Voltage-Gated Sodium Channels

Sinomenine is a pure alkaloid with immunosuppressive effects that is extracted from the Chinese medicinal plant Sinomenium acutum. We studied the therapeutic effects of sinomenine on inflammatory bowel disease. In this study, we randomly divided mice into the following ten groups: Control group; DSS-induced colitis group; Salicylazosulfapyridine (SASP)-treated group; Chitosan-treated group; low-, medium-, and high-dose sinomenine-treated and sinomenine enteric-coated microspheres-treated groups. We recorded changes in colon length, disease activity index (DAI), and colon pathology, measured TLR4, MyD88, SIGIRR, NF-κB p65 protein levels and inflammatory serum cytokine levels. Except for the Control group, the weight of mice in each group decreased, the DAI of the DSS-induced colitis group was significantly higher than the other groups, and the DAIs of the sinomenine- and sinomenine enteric-coated microspheres-treated groups were significantly lower than that of the SASP-treated group. TLR4, MyD88, NF-κB p65 and proinflammatory cytokine expressions decreased dose dependently in the sinomenine and sinomenine enteric-coated microspheres-treated groups and were generally lower in the sinomenine enteric-coated microspheres groups. However, SIGIRR and anti-inflammatory IL-10 expressions exhibited the opposite pattern. Based on the superior therapeutic effect, sinomenine enteric-coated microspheres might regulate TLR/NF-κB signaling and would be beneficial for an effective and safe therapy of inflammatory bowel disease.

Topics: Animals; Anti-Inflammatory Agents; Colitis; Colon; Dextran Sulfate; Disease Models, Animal; Disease Progression; Down-Regulation; Drug Delivery Systems; Female; Inflammatory Bowel Diseases; Medicine, Chinese Traditional; Mice; Mice, Inbred BALB C; Microspheres; Morphinans; Myeloid Differentiation Factor 88; NF-kappa B; Sinomenium; Toll-Like Receptor 4

Sinomenine hydrochloride (SH) has been investigated for its anti-tumor growth effect. We have previously reported that SH inhibited breast cancer cell proliferation via MAPKs signaling. However, whether SH could inhibit tumor metastasis has not been fully explored. In this study, we found that SH suppressed the metastasis potential of breast cancer cells. The wound healing and transwell assays showed that SH inhibited the migration and invasion ability of both 4T1 and MDA-MB-231 breast cancer cells. The orthotopic mouse model of 4T1 and the experimental mouse model of MDA-MB-231-luc (MDA-MB-231 cell line expressing firefly luciferase) demonstrated that SH treatment inhibited breast cancer metastasis by inhibiting epithelial-mesenchymal transition (EMT) and cancer stem cell (CSC) properties without obvious hepatotoxicity and renal toxicity. We also found that SH decreased spleen volume and weight in both mouse models, especially in the 4T1 mouse model. IL-6, a strong inflammatory factor causing EMT, was remarkably reduced. Overall, this anti-metastasis effect of SH could be possibly caused by attenuating inflammatory reaction, which led to inhibition of EMT and CSC characteristics of breast cancer cells. This study, together with our previous one, provides more evidence of SH as a potential drug for breast cancer therapy.

Topics: Animals; Antirheumatic Agents; Cell Line, Tumor; Disease Models, Animal; Epithelial-Mesenchymal Transition; Female; Humans; Inflammatory Breast Neoplasms; Mammary Neoplasms, Experimental; Mice; Morphinans; Neoplasm Metastasis; Neoplastic Stem Cells; RAW 264.7 Cells

Sinomenine is originally derived from medicinal herb and used preferentially in treatment of rheumatoid diseases in Far East regions. SIN has strong anti-inflammatory and immune-regulatory properties, acting mainly through inhibiting NF-kB signaling. Although the upstream target through which SIN affects NF-kB activity is unknown, evidence suggests that SIN might regulate inflammation through Nrf2 signaling. In this study we explored the role of Nrf2 in mediating SIN's anti-inflammation and kidney protection in a mouse model of obstructive nephropathy. We found that SIN is an activator of Nrf2 signaling. It markedly increased Nrf2 protein level, Nrf2 nuclear translocation, Nef2 transcription capacity, and the downstream protein expression. We further demonstrated that SIN activation of Nrf2 is likely due to its repression of the Nrf2 inhibitor Keap1 since it drastically reduced Keap1 protein through the PKC-sensitive ubiquitination-proteasomal degradation. SIN treatment of nephropathy mice effectively reduced the kidney damage and inflammatory responses, balanced renal oxidative stress, and improved the pathological protein expression in an Nrf2 dependent manner. In addition, SIN also Nrf2-dependently modulated macrophage M1/M2 polarization and inhibited the IkBα phosphorylation and NF-kB nuclear translocation, hence revealing an important upstream event that contributed to its anti-inflammation and tissue protection. Taken together our study has identified a novel pathway through which SIN exerts its anti-inflammation and renal protective functions, and provided a molecular basis for SIN potential applications in the treatment of kidney and other inflammatory disorders.

Topics: Acute Kidney Injury; Animals; Disease Models, Animal; Heme Oxygenase-1; Humans; Inflammation; Kidney; Macrophages; Mice; Morphinans; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress; Signal Transduction

The objective of this study is to investigate the therapeutic effect of sinomenine (SIN) on rat cerebral ischemia-reperfusion (IR) injury and the molecular mechanism.. One hundred thirty-five rats were equally randomized into sham-operated group, middle cerebral artery occlusion (MCAO) group, and SIN group, and reversible rat MCAO model was made according to the Longa method for the MCAO and SIN groups. Then, 15 rats from each group were decapitated at 6, 12, and 24 hours after reperfusion to obtain brain tissue samples. Rats in the SIN group were injected with sinomenine by tail vein (90 mg/kg) 1 hour before ischemia; rats in the MCAO and sham-operated groups were administrated with the same volume of saline. Neurological severity score (NSS), infarction volume, ischemic brain water content, and blood-brain barrier (BBB) permeability were determined at corresponding time points. Acid-sensing ion channel (ASIC) 1a mRNA level was determined by quantitative real-time polymerase chain reaction; ischemic brain contents of lactic acid (LD), lactic dehydrogenase (LDH), ATPase, and inflammatory factors were determined by spectrophotometric method.. At 12 hours after reperfusion and since then, NSS in the SIN group decreased obviously; infarction volume, brain water content, and BBB permeability in the SIN group were lower than those in the MCAO group (P < .05). IR injury resulted in the upregulation of the contents of ASIC1a mRNA, LD, LDH, and inflammatory factors and the downregulation of the contents of ATPase, while SIN could reverse the upregulation/downregulation effect induced by IR injury (P < .05).. Through its anti-inflammation effect, which alleviates acidosis, improves energy metabolism, and inhibits ASIC1a level, SIN protects ischemic rat brain against cerebral IR injury.

Topics: Acid Sensing Ion Channels; Acidosis; Animals; Anti-Inflammatory Agents; Blood-Brain Barrier; Brain; Capillary Permeability; Cytoprotection; Disease Models, Animal; Energy Metabolism; Infarction, Middle Cerebral Artery; Inflammation Mediators; Male; Morphinans; Neuroprotective Agents; Rats, Sprague-Dawley; Reperfusion; Reperfusion Injury; Time Factors

Astrocyte-mediated neuroinflammation plays a critical role in ischemic stroke-induced secondary cerebral injury. Previous studies have suggested that the dopamine D2 receptor (DRD2) acts as a key target in regulating the neuroinflammatory response. However, the underlying molecular mechanisms are still unknown, and effective DRD2 agonists are lacking. In the present study, we examined the anti-inflammatory and neuroprotective effects of sinomenine (Sino), a monomeric compound with potential immunoregulatory properties in nervous system.. TTC staining, apoptosis assay, evaluation of brain edema, and neurological assessment were performed in the middle cerebral artery occlusion (MCAO) mouse model. Primary astrocytes exposed to oxygen glucose deprivation (OGD) were used in the in vitro experiments. Quantitative PCR was applied to assess the levels of inflammatory cytokines. Multi-labeling immunofluorescence, Western blot, co-immunoprecipitation, and electrophoretic mobility shift assay (EMSA) were also used to investigate the molecular mechanisms underlying the Sino-mediated anti-inflammatory effects in vivo and in vitro.. Sino remarkably attenuated the cerebral infarction and neuronal apoptosis, reduced the levels of inflammatory cytokines, and alleviated neurological deficiency in MCAO mice. Sino significantly inhibited astrocytic activation and STAT3 phosphorylation as well as increased DRD2 and αB-crystallin (CRYAB) expression after MCAO. In vitro, Sino blocked OGD-induced activation of STAT3 and generation of pro-inflammatory cytokines in primary astrocytes, and these effects were significantly abolished by either DRD2 or CRYAB knockdown. Additionally, Sino induced up-regulation and nuclear translocation of CRYAB in astrocytes and enhanced the interaction between CRYAB and STAT3, which further inhibited the activation and DNA-binding activity of STAT3.. Our study demonstrates that Sino activates astrocytic DRD2 and thereby suppresses neuroinflammation via the CRYAB/STAT3 pathway, which sheds some light on a promising therapeutic strategy for ischemic stroke.

Topics: alpha-Crystallin B Chain; Animals; Animals, Newborn; Antirheumatic Agents; Astrocytes; Brain Edema; Cells, Cultured; Cerebral Infarction; Disease Models, Animal; Encephalitis; Hypoxia; Infarction, Middle Cerebral Artery; Mice; Mice, Inbred C57BL; Morphinans; Nervous System Diseases; Receptors, Dopamine D2; RNA Interference; Signal Transduction; STAT3 Transcription Factor

Background and aims We have previously reported that sinomenine, an alkaloid isolated from the root of the plant Sinomenium acutum, had antinociceptive effect in rodent models of acute inflammatory or neuropathic pain. As a traditional medicine, sinomenine is used in China to treat rheumatoid arthritis (RA). Methods In the present study, we evaluated the potential antinociceptive effect of sinomenine in a mouse model of RA, collagen type II antibody (CII Ab) induced arthritis (CAIA) after acute and chronic administration. Results As single administration, sinomenine at 40 or 80 mg/kg significantly reduced mechanical hypersensitivity both at the time of peak joint inflammation (days 11-19 after CII Ab injection) or during the post-inflammatory phase (days 35-54). No tolerance to the effect of 80 mg/kg sinomenine was observed during repeated injection twice a day for 5 days from day 11 to day 19 or from day 49 to day 53 after CII Ab injection in CAIA mice. Conclusions We have shown that sinomenine is effective in alleviating localized and spread hypersensitivities in CAIA mice both during acute inflammation and in post-inflammatory phase. Further, repeated sinomenine administration has elevated the baseline mechanical threshold without producing tolerance. Implications Sinomenine may be clinically useful to treat chronic pain in RA, including wide-spread pain which appears to be a difficult clinical problem despite the improvement in the acute treatment of RA by disease modifying agents.

Topics: Animals; Antirheumatic Agents; Arthritis, Rheumatoid; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Hyperalgesia; Mice, Inbred CBA; Morphinans; Time Factors; Touch

Sepsis, a systemic inflammatory response to infection, is the major cause of death in intensive care units (ICUs). The mortality rate of sepsis remains high even though the treatment and understanding of sepsis both continue to improve. Sinomenine (SIN) is a natural alkaloid extracted from Chinese medicinal plant Sinomenium acutum, and its hydrochloride salt (Sinomenine hydrochloride, SIN-HCl) is widely used to treat rheumatoid arthritis (RA). However, its role in sepsis remains unclear. In the present study, we investigated the role of SIN-HCl in sepsis induced by cecal ligation and puncture (CLP) in BALB/c mice and the corresponding mechanism. SIN-HCl treatment improved the survival of BALB/c mice that were subjected to CLP and reduced multiple organ dysfunction and the release of systemic inflammatory mediators. Autophagy activities were examined using Western blotting. The results showed that CLP-induced autophagy was elevated, and SIN-HCl treatment further strengthened the autophagy activity. Autophagy blocker 3-methyladenine (3-MA) was used to investigate the mechanism of SIN-HCl in vitro. Autophagy activities were determined by examining the autophagosome formation, which was shown as microtubule-associated protein light chain 3 (LC3) puncta with green immunofluorescence. SIN-HCl reduced lipopolysaccharide (LPS)-induced inflammatory cytokine release and increased autophagy in peritoneal macrophages (PM). 3-MA significantly decreased autophagosome formation induced by LPS and SIN-HCl. The decrease of inflammatory cytokines caused by SIN-HCl was partially aggravated by 3-MA treatment. Taken together, our results indicated that SIN-HCl could improve survival, reduce organ damage, and attenuate the release of inflammatory cytokines induced by CLP, at least in part through regulating autophagy activities.

Topics: Adenine; Animals; Autophagy; Cecum; Cell Line; Cytokines; Disease Models, Animal; Kidney; Lipopolysaccharides; Liver; Lung; Macrophages; Male; Mice; Mice, Inbred BALB C; Microtubule-Associated Proteins; Morphinans; Sepsis; Survival Rate

Rheumatoid arthritis (RA) is a chronic, systemic inflammatory disorder that affects about 1% of the population worldwide. RA is mainly manifested by persistent synovitis and progressive joint destruction. The aim of the present study was to examine the anti-arthritis effects of SND-117, a sinomenine bivalent that is obtained from the structure modification of a clinically available anti-RA drug, sinomenine. The arthritis model (CIA) was established by immunizing DBA/1 mice with type II collagen, and the arthritis scores including inflammation, joint destruction and bone erosion were assessed after booster immunization for 3weeks. The levels of cytokines such as IL-1β, IL-6 and TNF-α were analyzed by quantitative PCR and ELISA. The TNF-α induced NF-κB activation in fibroblast-like synovial cells (FLSCs) was analyzed by Western blot. SND-117 significantly relieved the inflammatory symptoms of collagen-induced arthritis, reduced bone erosion and joint destruction in CIA mice. The serum levels of IL-1β, IL-6 and TNF-α of CIA mice were markedly decreased by SND-117. SND-117 also strongly inhibited the phosphorylation and nuclear translocation of NF-κB p65 in FLSCs upon TNF-α stimulation. These data demonstrated that SND-117 could effectively block the pathogenesis of collagen-induced arthritis in CIA mice via inhibition of NF-κB signaling, and might provide potential clinic benefits in rheumatoid arthritis management.

Topics: Animals; Antirheumatic Agents; Arthritis, Rheumatoid; Bone and Bones; Collagen Type II; Cytokines; Disease Models, Animal; Fibroblasts; Humans; Immunization, Secondary; Inflammation Mediators; Male; Mice; Mice, Inbred DBA; Morphinans; NF-kappa B; Sinomenium; Synovitis

To explore the role of sinomenine in treatment of allergic rhinitis mice model and its possible mechanism.. We used ovalbumin (OVA) to make allergic rhinitis model of BALB/c mice. Saline was used in the control group. When we challenged the mice with OVA intranasally, the mice in sinomenine treatment group were feed by the food containing sinomenine. Mice were then killed 24 h after the last OVA challenge. The noses of mice from each group were removed en bloc and fixed, then each section was stained with hematoxylin and eosin. ELISA assay was used to measure the concentration of anti-OVA IgE, IL-4 and IFN-gamma. The proteins expressive level of T-bet and GATA3 were examined.. Nasal mucosa of the mice in sinomenine treatment group were not hyperplasia and without obvious infiltration of eosinophils. The concentration of anti-OVA IgE, IL-4 and IFN-gamma in the serum and T-bet and GATA3 expression levels of sinomenine treatment group were lower than those of allergic rhinitis group.. The sinomenine can be used to treat allergic rhinitis mice, and the mechanism may rely on the improvements of the Th1/Th2 imbalance.

Topics: Animals; Disease Models, Animal; Eosinophils; GATA3 Transcription Factor; Immunoglobulin E; Interferon-gamma; Interleukin-4; Male; Mice; Mice, Inbred BALB C; Morphinans; Nasal Mucosa; Phytotherapy; Rhinitis, Allergic; Rhinitis, Allergic, Perennial; T-Box Domain Proteins; Th1 Cells; Th2 Cells

To observe the analgesic effect of sinomenine on the neuropathic pain rat model induced by SSNI, and discuss its impact on monoamine neurotransmitters in striatal extracellular fluid.. Male SD rats were randomly divided into the sham operation group, the SSNI model group, the gabapentin group (100 mg x kg(-1)), the sinomenine high dose group (40 mg x kg(-1)) and the sinomenine low dose group (20 mg x kg(-1)). Mechanical hyperalgesia and cold pain sensitivity were evaluated by Von Frey hairs and cold spray. Striatum was sampled by microdialysis. High performance liquid chromatography-electrochemical detector (HPLC-ECD) were used to detect the content of such neurotransmitters as monoamine neurotransmitters noradrenaline (NE), dopamine (DA), 5-hydroxy tryptamine (5-HT) and their metabolites dihydroxyphenylacetic phenylacetic acid (DOPAC) and 5-hydroxyindoleacetic acid (5-HIAA).. SSNI model rats showed significant improvement in mechanical withdrawal threshold and cold pain sensitivity, significant decrease in intracerebral NE and notable increase in DA, 5-HT and their metabolites. Compared with the model group, the sinomenine high dose group showed significant increase in mechanical withdrawal threshold at 60, 90, 180 and 240 min after abdominal administration (P < 0.01), significant decrease in cold pain sensitivity score during 30-240 min (P < 0.05). Sinomenine can significantly up-regulated NE content in striatal extracellular fluid during 45-135 min (P < 0.05), remarkably reduce DA content and DOPAC at 45, 75 and 135 min (P < 0.05), 5-HT content during 45-135 min, DOPAC during 75-165 min (P < 0.05), and 5-HIAA during 45-135 min (P < 0.05).. Sinomenine has the intervention effect on neuropathic pain in SSNI model rats. Its mechanism may be related to disorder of monoamine neurotransmitters in striatal extracellular fluid.

Topics: Analgesics; Animals; Biogenic Monoamines; Disease Models, Animal; Extracellular Fluid; Male; Morphinans; Neostriatum; Neurotransmitter Agents; Rats; Rats, Sprague-Dawley; Sciatic Nerve

Sinomenine (SIN) is a purified alkaloid from the Chinese herb Sinomenium acutum. Previous studies demonstrated that SIN possesses anti-inflammatory and anti-apoptotic properties. We thus in the present report conducted studies to examine its impact on ischemia reperfusion (IR) induced renal injury. Precondition of mice with 200 mg/kg of SIN provided significant protection for mice against IR-induced renal injury as manifested by the attenuated serum creatinine (Cre) and blood urea nitrogen (BUN) along with less severity for histological changes and tubular cell apoptosis. In line with these results, treatment of mice with SIN suppressed IR-induced inflammatory infiltration and the expression of chemokine CXCL-10, adhesion molecule ICAM-1, and cytokines TNF-а/IL-6. Mechanistic studies revealed that SIN inhibits NF-κB transcriptional activity to suppress IR-induced inflammatory response in the kidney, while it attenuates MAP kinase signaling to prevent tubular cells undergoing apoptosis after IR insult. Altogether, our data support that SIN could be a useful therapeutic agent for prevention and treatment of IR-induced renal injury in the clinical settings.

Topics: Animals; Anti-Inflammatory Agents; Apoptosis; Biomarkers; Blood Urea Nitrogen; Chemokine CXCL10; Creatinine; Cytoprotection; Disease Models, Animal; Inflammation Mediators; Intercellular Adhesion Molecule-1; Interleukin-6; Kidney Tubules; Male; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Morphinans; Nephritis; NF-kappa B; Reperfusion Injury; Time Factors; Tumor Necrosis Factor-alpha

Sinomenine (SN), a bioactive alkaloid, has been utilized clinically to treat rheumatoid arthritis in China. Our preliminary experiments indicated that it could protect PC12 cells from oxygen-glucose deprivation-reperfusion (OGD-R), we thus investigated the possible effects of SN on cerebral ischaemia and the related mechanism.. Middle cerebral artery occlusion in rats was used as an animal model of ischaemic stroke in vivo. The mechanisms of the effects of SN were investigated in vitro using whole-cell patch-clamp recording, calcium imaging in PC12 cells and rat cortical neurons subjected to OGD-R.. Pretreatment with SN (10 and 30 mg·kg(-1) , i.p.) significantly decreased brain infarction and the overactivation of calcium-mediated events in rats subjected to 2 h ischaemia followed by 24 h reperfusion. Extracellular application of SN inhibited the currents mediated by acid-sensing ion channel 1a and L-type voltage-gated calcium channels, in the rat cultured neurons, in a concentration-dependent manner. These inhibitory effects contribute to the neuroprotection of SN against OGD-R and extracellular acidosis-induced cytotoxicity. More importantly, administration of SN (30 mg·kg(-1) , i.p.) at 1 and 2 h after cerebral ischaemia also decreased brain infarction and improved functional recovery.. SN exerts potent protective effects against ischaemic brain injury when administered before ischaemia or even after the injury. The inhibitory effects of SN on acid-sensing ion channel 1a and L-type calcium channels are involved in this neuroprotection.

Topics: Acid Sensing Ion Channels; Animals; Animals, Newborn; Blotting, Western; Brain Ischemia; Calcium Channels, L-Type; Cell Culture Techniques; CHO Cells; Cricetinae; Cricetulus; Cytoprotection; Disease Models, Animal; Glucose; Male; Morphinans; Nerve Tissue Proteins; Neurons; Neuroprotective Agents; Oxygen; Patch-Clamp Techniques; PC12 Cells; Rats; Rats, Sprague-Dawley; Sodium Channels

To observe the effect of alcohol extracts from orientivne and its effective component sinomenine on withdrawal syndromes and neurotransmitter of morphine-abstinent mice and on intracellular calcium level in morphine-dependent neuronal-cells line. To study the detoxification of alcohol extracts from orientvine and sinomenine on morphine-dependent animal and explore the mechanism of its effect.. The effect of alcohol extracts from orientivne and sinomenineon on abstinent syndromes was observed by experiment study on morphine-dependent ex vivo ileum from guinea pigs and morphine-dependent mice. The morphine-dependent model mice were established by injection on dosage increasing by degrees. The hypothalamic monomine neurotransmitters such as NA, DA, 5-HT were tested by fluorospectrophotometry. Morphine-dependent cell line was established by administering morphine at different doses into the culture medium. The cells were stained with fluo-3 and the intracellular calcium level was measured by flow cytometry.. Alcohol extracts from orientvine and sinomenine could alleviate withdrawal contractile response of morphine-dependent ex vivo ileum from guinea pigs and withdrawal syndromes of morphine-dependent mice, decrease the concentration of the neurotransmitters, and elevate the intracellular calcium level and inhibit the decreasing of Ca2+ induced by naloxone.. Alcohol extracts from orientvine and sinomenine have some effects on withdrawal syndromes which may be related to inhibiting neurotransmitters and the regulation of intracellular calcium concentration.

Topics: Animals; Calcium; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Ethanol; Female; Guinea Pigs; Ileum; Male; Mice; Morphinans; Morphine; Morphine Dependence; Muscle Contraction; Neurons; Norepinephrine; Random Allocation; Rats; Rats, Sprague-Dawley; Sinomenium; Substance Withdrawal Syndrome

To investigate the effects of intra-articular injection of sinomenine on interleukin-1beta (IL-1beta) content in synovial fluid and serum in rabbits with osteoarthritis (OA).. Degenerative osteoarthritis in the knee joint of left posterior limb was induced in 36 rabbits by full extension using plaster cast for 6 weeks. Then the rabbits were randomly divided into untreated group, hyaluronic acid group and sinomenine group. Another 4 normal rabbits were selected as normal control group. Rabbits in the sinomenine group and the hyaluronic acid group received intraarticular injections of sinomenine and hyaluronic acid once a week for 5 weeks, respectively. The content of IL-1beta in synovial fluid and serum were measured before and after treatment by enzyme-linked immunosorbent assay. The pathological changes of cartilaginous tissue were analyzed by using Mankin's score.. Compared with those in the untreated group, synovial fluid and serum IL-1beta contents in the sinomenine group and the hyaluronic acid group were significantly decreased (P<0.01). And the synovial fluid and serum IL-1beta contents in the sinomenine group were lower than those in the hyaluronic acid group (P<0.05). The mean Mankin's score of cartilage in the sinomenine group was significantly lower than that in hyaluronic acid group (P<0.05).. Sinomenine may reduce the degree of articular degeneration in rabbit with OA through decreasing the content of IL-1beta in synovial fluid and serum.

Topics: Animals; Disease Models, Animal; Hyaluronic Acid; Interleukin-1beta; Male; Morphinans; Osteoarthritis, Knee; Rabbits; Serum; Synovial Fluid

To evaluate the possible role of alkaloid sinomenine (SIN) on chronic rejection in rat heart transplantation model.. After a brief course of cyclosporine A (CsA), DA recipients of PVG hearts were treated with placebo, SIN, CsA, or a combination of both drugs. Grafts were analyzed morphometrically and by immuno-histochemistry. Expressions of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and endothelin 1 (ET-1) were assessed by reverse transcription-polymerase chain reaction.. Cardiac grafts of SIN-treated rats showed a mild degree of vasculopathy compared with untreated rats or CsA-treated recipients. Degree of vasculopathy was significantly reduced in rats treated with combined SIN and CsA than rats receiving either drug alone. Treatment with SIN alone did not affect gene expressions of bFGF, VEGF, and ET-1 while expressions of bFGF, VEGF, and ET-1 were significantly reduced by combined treatment with SIN and CsA.. These results demonstrated a potential value of SIN, in combination with low-dose CsA to attenuate the vasculopathy in this rat model of chronic cardiac allograft rejection.

Topics: Animals; Disease Models, Animal; Graft Rejection; Graft Survival; Heart Transplantation; Male; Morphinans; Phytotherapy; Rats

For exploring the mechanism of the anti-inflammatory and anti-rheumatic effect of sinomenine (SN), the actions of different dosage of SN were observed in vitro on the expression of cytokines, tumor necrosis factor (TNF-alpha) and interleukin-1beta (IL-1beta), as well as the activity of nuclear factor-kappaB (NF-kappaB) and the inhibitory kappaB-alpha (IkappaB-alpha) protein level of peritoneal macrophages (PMs) and synoviocytes in adjuvant arthritis (AA) rats. In this study, the experimental rat model of AA was used and PMs and synoviocytes were collected. The mRNAs of TNF-alpha and IL-1beta were detected with reverse transcription-polymerase chain reaction (RT-PCR) and NF-kappaB activity was measured by electrophoretic mobility shift assay (EMSA). The IkappaB-alpha protein level in the cytoplasma was detected by Western blot. Our results showed that expression of mRNAs of TNF-alpha and IL-1beta and NF-kappaB activity by PMs and synoviocytes were markedly increased compared to control group (P<0.05). In a definite concentration ranging from 30 to 120 microg/ml, SN showed inhibiting effect on the NF-kappaB activity and the expression of the mRNAs of TNF-alpha and IL-1beta in AA rats in a concentration-dependent manner (P<0.05). Positive correlations were found between changes of NF-kappaB activity and expression of TNF-alpha mRNA and IL-1beta mRNA (P<0.01). IkappaB-alpha protein level was increased by various dosages of SN in comparison with control group (P<0.01). In conclusion, SN decreases the mRNA expression of TNF-alpha and IL-1beta by inhibiting the NF-kappaB binding activity, which is mediated through up-regulating the IkappaB-alpha expression of PMs and synoviocytes in AA rats.

Topics: Animals; Arthritis, Experimental; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; I-kappa B Proteins; Interleukin-1; Macrophages, Peritoneal; Male; Morphinans; NF-kappa B; Plants, Medicinal; Rats; Rats, Wistar; RNA, Messenger; Synovial Membrane; Tumor Necrosis Factor-alpha; Up-Regulation

The present study was designed to investigate the putative anxiolytic-like effect of sinomenine in three experimental models of anxiety in male rats and mice. Use of the elevated plus-maze test revealed that sinomenine (20 and 40 mg/kg, p.o.) increased the percentage of open arm entries and diazepam (2 mg/kg, p.o.) increased the percentage of open arm entries, the percentage of time spent on open arms and total arm entries in mice. In the light/dark transition test, sinomenine (20 and 40 mg/kg, p.o.) increased time spent in the light area and diazepam (2 mg/kg, p.o.) increased time spent in the light area and the overall movements in mice. In the social interaction test, the sinomenine-treated animals significantly increased social interaction time in low light unfamiliar (7 mg/kg, p.o.) and high light unfamiliar conditions (7 and 14 mg/kg, p.o.) as well as diazepam (3 mg/kg, p.o.). Sinomenine (28 mg/kg, p.o.) can also decrease squares entered in rats in social interaction test under low light unfamiliar condition. In the open-field test, sinomenine (160 mg/kg) decreased squares entered in mice. Thus, these findings indicated that sinomenine exhibited anxiolytic-like effect.

Topics: Animals; Anti-Anxiety Agents; Anxiety; Behavior, Animal; Darkness; Disease Models, Animal; Dose-Response Relationship, Drug; Light; Male; Maze Learning; Mice; Morphinans; Rats; Rats, Wistar; Social Behavior; Species Specificity

The effects of treatment with sinomenine, a pure alkaloid extracted from the chinese medical plant Sinomenium acutum, were investigated in rat adjuvant arthritis (AA) and antigen-induced arthritis (AIA). In AA, long-term, intraperitoneal (i.p.) treatment induced significant improvement of arthritic score, hind paw swelling, body weight and erythrocyte sedimentation rate (ESR) beginning past the clinical peak of the disease. In-acute AIA, short and middle-term treatment with sinomenine around and following induction of arthritis induced a dose-dependent decrease of both joint swelling and ESR, starting after the peak of arthritis, and a significant reduction of joint destruction on day 3. There was no rebound of the arthritic signs following suspension of treatment. Long-term treatment of chronic AIA partially ameliorated clinical parameters and significantly counteracted joint destruction. Maximal plasma concentrations of 22.5 micrograms/ml, fast wash out (half-life 4.24 +/- 0.99 h; mean +/- S.E.M.) and no evidence of accumulation of sinomenine were observed following single or repeated i.p. injection of 150 mg/kg. In vitro, sinomenine markedly inhibited proliferation of synovial fibroblasts from AIA or normal rats, both at rest and following activation with either transforming growth factor beta 2 (TGF-beta 2) or interleukin-1 beta (IL-1 beta). The effect was dose-dependent and half-maximal inhibition of proliferation occurred at 20.6 micrograms/ml, that is, within the in vivo therapeutic range of the drug. Late therapeutic effects of sinomenine in rat arthritic models despite early start of treatment may be related to its antiproliferative effects on synovial fibroblasts in addition to its previously reported anti-inflammatory properties.

Topics: Adjuvants, Immunologic; Animals; Anti-Inflammatory Agents, Non-Steroidal; Antigens; Arthritis, Experimental; Cell Division; Cells, Cultured; Cytokines; Disease Models, Animal; Female; Fibroblasts; Morphinans; Rats; Rats, Inbred Lew; Stimulation, Chemical; Synovial Membrane